The Role of Yes Associated Protein (YAP) in Hypertrophic Cardiomyopathy

Yes 相关蛋白 (YAP) 在肥厚性心肌病中的作用

基本信息

批准号：
10389312
负责人：
Orlando Chirikian
金额：
$ 3.9万
依托单位：
UNIVERSITY OF CALIFORNIA SANTA BARBARA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10389312
关键词：
Actins Address Affect Biochemical Biomechanics Cardiac Cardiac Myocytes Cardiomyopathies Cardiovascular Diseases Cell Nucleus Cells Clinical Cytoskeleton Desmin Development Disease Elements Environment Event Exhibits Fibrosis Generations Growth Heart Heart Diseases Homeostasis Human Hypertrophic Cardiomyopathy Hypertrophy In Vitro Individual Inherited Kinetics Knowledge Label Lead Measures Mechanical Stress Mechanics Mediating Microtubules Modeling Molecular Morphology Mutation Myosin ATPase Nuclear Nuclear Protein Organ Organ Size Pathologic Pathway interactions Pharmaceutical Preparations Phenotype Point Mutation Production Protein Dynamics Proteins Regulation Research Research Training Role Signal Transduction Signaling Protein Testing Therapeutic Traction Troponin disease phenotype extracellular genetic regulatory protein induced pluripotent stem cell derived cardiomyocytes insight mechanical signal mutant myosin-binding protein C non-muscle myosin heavy chain-B novel optogenetics response tool transcriptome transmission process

项目摘要

Project Summary/Abstract Hypertrophy Cardiomyopathy (HCM) is the most prevalent hereditary cardiovascular disease – affecting 1 in 500 individuals. Advanced forms of the disease clinically present with hypercontractility, hypertrophy (enlargement of the organ and individual cardiomyocytes) and fibrosis. Several single-point mutations in b- myosin heavy chain (MYH7), Myosin Binding Protein C (MYBPC3), and Troponin (cTn) have been associated with HCM and increased contractility at the organ level. However, the kinetics at the molecular level remain unclear, as different sarcomeric protein mutations can result in increased, decreased, or unchanged force production. A knowledge gap persist in understanding how these altered kinetics at the molecular level lead to the more advanced hypertrophic phenotype of HCM at the cellular level. Interestingly, the Hippo Pathway has been demonstrated to be activated during developmental growth, quiescent during cardiac homeostasis, and reactivated in pathological growth (i.e. HCM). However its involvement in the disease, in particular the initiation of the hypertrophic phenotype, is poorly understood. Here, we aim to understand whether homeostatic mechanical signaling through the canonical growth regulator, Hippo-YAP, is altered 1) by changes in the biomechanics of single HCM mutant cardiomyocytes and 2) by alterations in the mechanical environment. We propose to use human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) genetically edited to harbor point mutations associated with HCM, as a reduce ordered model to study the relationship between mechanical signaling and hypertrophic growth. We will modulate mechanical stresses (i.e. diseased conditions) in healthy and diseased cardiomyocytes by treatment with inotropic drugs and culture in fibrotic-like stiff conditions and track the resulting signaling events by fluorescently labeling the key regulatory protein of the Hippo pathway (YAP). To further elucidate the mechanism by which YAP is contributing to the phenotypes of HCM we have developed a novel optogenetic tool, termed OptoYAP, which provides full temporal and spatial control of the Hippo pathway. Lastly we aim to understand the mechanism behind the reactivation of YAP in pathological conditions by perturbing the mechanical signaling by the nucleus. We hypothesize that 1) changes in force production alter the homeostatic mechano-signaling of the Hippo pathway to initiate cellular hypertrophy and 2) subsequent changes to the extracellular environment (stiffening) compounds this effect leading to a feedforward signal progressing the disease phenotypes. 3) pathological YAP signaling is driven by excessive force transmission by the cytoskeleton resulting in nuclear deformation. Our results will provide insights into HCM progression and provide a testbed for therapeutic options in treating HCM.

项目概要/摘要肥厚型心肌病 (HCM) 是最常见的遗传性心血管疾病 - 影响 1 % 500 名晚期患者临床表现为收缩过度、肥大。（器官和单个心肌细胞增大）和纤维化。b-中的几个单点突变。肌球蛋白重链 (MYH7)、肌球蛋白结合蛋白 C (MYBPC3) 和肌钙蛋白 (cTn) 相关然而，分子水平上的动力学仍然存在。尚不清楚，因为不同的肌节蛋白突变可能导致力量增加、减少或不变在理解这些分子水平上的动力学改变如何导致生产方面仍然存在知识差距。 HCM 在细胞水平上更先进的肥厚表型。已被证明在发育生长期间被激活，在心脏稳态期间处于静止状态，并且在病理性生长（即 HCM）中重新激活，但它参与了疾病，特别是起始。对于肥大表型的影响，人们知之甚少。在这里，我们的目的是了解体内平衡是否存在。通过经典生长调节剂 Hippo-YAP 的机械信号传导会因 1) 的变化而改变单个 HCM 突变心肌细胞的生物力学和 2) 通过机械环境的改变。提议使用经过基因编辑的人类诱导多能干细胞衍生的心肌细胞（hiPSC-CM）含有与 HCM 相关的点突变，作为研究之间关系的降序模型我们将调节机械应力（即疾病状况）。通过用正性肌力药物治疗和在纤维化样硬块中培养，在健康和患病的心肌细胞中产生条件并通过荧光标记关键调节蛋白来跟踪由此产生的信号事件 Hippo 通路 (YAP) 进一步阐明 YAP 影响表型的机制。 HCM 我们开发了一种新型光遗传学工具，称为 OptoYAP，它提供了完整的时空信息最后，我们的目标是了解 YAP 重新激活背后的机制。我们通过扰乱细胞核的机械信号来调节病理状况：1）变化。强制生产改变 Hippo 通路的稳态机械信号以启动细胞肥大和 2) 随后细胞外环境的变化（硬化）加剧了这种效应导致前馈信号进展疾病表型 3) 病理性 YAP 信号传导由以下因素驱动。我们的结果将提供细胞骨架过度的力传递导致核变形。深入了解 HCM 进展，并为治疗 HCM 的治疗方案提供试验台。